Buffered microencapsulated compositions and methods

ABSTRACT

A microcapsule composition comprising at least one polymer substantially disposed as a semipermeable shell around a buffered solution and at least one therapeutic agent, wherein the therapeutic agent permeates the shell, and wherein the composition is suitable for topical epithelial cells of mammal.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.16/207,515 filed Dec. 3, 2018, which is a continuation of U.S. patentapplication Ser. No. 15/921,269 filed Mar. 14, 2018, which is adivisional of U.S. patent application Ser. No. 15/791,554 filed Oct. 24,2017, which is a divisional of U.S. patent application Ser. No.13/619,128 filed Sep. 14, 2012, which is a continuation-in-part of U.S.patent application Ser. No. 12/768,696, filed Apr. 27, 2010, whichclaims priority of U.S. Provisional Application No. 61/172,939, filedApr. 27, 2009, the contents of which are incorporated herein byreference in their entirety.

BACKGROUND OF THE INVENTION

The present invention is directed to compositions, compounds and methodsfor topical use of therapeutic agents on epithelial tissues. Variousconnective and epithelial tissue or tissues include skin, teeth, bone,and various connective tissues such as collagen, cartilage, tendons,ligaments and other dense connective tissue and reticular fibers (thatcontains type III collagen) of a mammal, including a human being.

Calcium phosphates are a class of minerals containing, but not limitedto, calcium ions together with orthophosphates, metaphosphates and/orpyrophosphates that may or may not contain hydrogen or hydroxide ions.

For purposes of definition in this specification, “remineralization” isthe process of restoring minerals, in the form of mineral ions, to thehydroxyapatite latticework structure of a tooth. As used herein, theterm “remineralization” includes mineralization, calcification,recalcification and fluoridation as well as other processes by whichvarious particular ions are mineralized to the tooth. The term “teeth”or “tooth” as used herein includes the dentin, enamel, pulp and cementumof a tooth within the oral cavity of an animal, including a human being.

In certain embodiments, the present invention provides methods forwhitening the surface of a tooth material by using the compositions ofthe invention. For purposes of definition in this specification, asreferred to herein, a “tooth material” refers to natural teeth,dentures, dental plates, fillings, caps, crowns, bridges, dentalimplants, and the like, and any other hard surfaced dental prosthesiseither permanently or temporarily fixed to a tooth within the oralcavity of an animal, including a human being. As used herein, the terms“whitening” and “tooth whitening” used interchangeably, refer to achange in the visual appearance of a tooth as defined herein, preferablysuch that the tooth has a brighter shade or luster.

Conditions of the Bone

No currently practiced therapeutic strategy involves methods orcompositions that sufficiently stimulate or enhance the growth of newbone mass. The present invention provides compositions, products andmethods which serve to increase bone mineralization at localized sitesor remineralization of teeth directly in the oral cavity, and thus maybe utilized in conjunction with treatments of a wide variety ofconditions where it is desired to increase bone or tissue mass as aresult of any condition which can be improved by bioavailability ofphysiological salts, particularly of calcium and phosphate.

Certain changes in bone mass occur over the life span of an individual.After about the age of 40 and continuing to the last stages of life,slow bone loss occurs in both men and women. Loss of bone mineralcontent can be caused by a variety of conditions, and may result insignificant medical problems. If the process of tissue mineralization isnot properly regulated, the result can be too little of the mineral ortoo much—either of which can compromise bone health, hardness andstrength. A number of bone growth disorders are known which cause animbalance in the bone remodeling cycle. Chief among these are metabolicbone diseases such as osteoporosis, osteoplasia (osteomalacia), chronicrenal failure and hyperparathyroidism, which result in abnormal orexcessive loss of bone mass known as osteopenia. Other bone diseases,such as Paget's disease, also cause excessive loss of bone mass atlocalized sites.

Osteoporosis is a structural deterioration of the skeleton caused byloss of bone mass resulting from an imbalance in bone formation, boneresorption, or both. Bone resorption is the process by which osteoclastsbreak down bone and release the minerals, resulting in a transfer ofcalcium from bone fluid to the blood. Bone resorption dominates the boneformation phase, thereby reducing the weight-bearing capacity of theaffected bone. In a healthy adult, the rate at which bone is formed andresorbed is tightly coordinated so as to maintain the renewal ofskeletal bone. However, in osteoporotic individuals, an imbalance inthese bone remodeling cycles develops which results in both loss of bonemass and in formation of microarchitectural defects in the continuity ofthe skeleton. These skeletal defects, created by perturbation in theremodeling sequence, accumulate and finally reach a point at which thestructural integrity of the skeleton is severely compromised and bonefracture is likely. Although this imbalance occurs gradually in mostindividuals as they age, it is much more severe and occurs at a rapidrate in postmenopausal women. In addition, osteoporosis also may resultfrom nutritional and endocrine imbalances, hereditary disorders and anumber of malignant transformations.

Osteoporosis in humans is preceded by clinical osteopenia (bone mineraldensity that is greater than one standard deviation but less than 2.5standard deviations below the mean value for young adult bone), acondition found in approximately 25 million people in the United States.Another 7-8 million patients in the United States have been diagnosedwith clinical osteoporosis (defined as bone mineral content greater than2.5 standard deviations below that of mature young adult bone).Osteoporosis is one of the most expensive diseases for the health caresystem, costing billions of dollars annually in the United States. Inaddition to health care related costs, long-term residential care andlost working days add to the financial and social costs of this disease.Worldwide, approximately 75 million people are at risk for osteoporosis.

The frequency of osteoporosis in the human population increases withage, and among Caucasians is predominant in women, who compriseapproximately 80% of the osteoporosis patient pool in the United States.In addition in women, another phase of bone loss occurs possibly due topostmenopausal estrogen deficiencies. During this phase of bone loss,women can lose an additional 10% in the cortical bone and 25% from thetrabecular compartment. The increased fragility and susceptibility tofracture of skeletal bone in the aged is aggravated by the greater riskof accidental falls in this population. More than 1.5 millionosteoporosis-related bone fractures are reported in the United Stateseach year. Fractured hips, wrists, and vertebrae are among the mostcommon injuries associated with osteoporosis. Hip fractures inparticular are extremely uncomfortable and expensive for the patient,and for women correlate with high rates of mortality and morbidity.

Patients suffering from chronic renal (kidney) failure almostuniversally suffer loss of skeletal bone mass, termed renalosteodystrophy. While it is known that kidney malfunction causes acalcium and phosphate imbalance in the blood, to date replenishment ofcalcium and phosphate by dialysis does not significantly inhibitosteodystrophy in patients suffering from chronic renal failure. Inadults, osteodystrophic symptoms often are a significant cause ofmorbidity. In children, renal failure often results in a failure togrow, due to the failure to maintain and/or to increase bone mass.

Osteoplasia, also known as osteomalacia (“soft bones”), is a defect inbone mineralization (e.g., incomplete mineralization), and classicallyis related to vitamin D deficiency (1,25-dihydroxy vitamin D₃). Thedefect can cause compression fractures in bone, and a decrease in bonemass, as well as extended zones of hypertrophy and proliferativecartilage in place of bone tissue. The deficiency may result from anutritional deficiency (e.g., rickets in children), malabsorption ofvitamin D or calcium, and/or impaired metabolism of the vitamin.

Hyperparathyroidism (overproduction of the parathyroid hormone) is knownto cause malabsorption of calcium, leading to abnormal bone loss. Inchildren, hyperparathyroidism can inhibit growth, in adults the skeletonintegrity is compromised and fracture of the ribs and vertebrae arecharacteristic. The parathyroid hormone imbalance typically may resultfrom thyroid adenomas or gland hyperplasia, or may result from prolongedpharmacological use of a steroid. Secondary hyperparathyroidism also mayresult from renal osteodystrophy. In the early stages of the disease,osteoclasts are stimulated to resorb bone in response to the excesshormone present. As the disease progresses, the trabecular boneultimately is resorbed and marrow is replaced with fibrosis, macrophagesand areas of hemorrhage as a consequence of microfractures, a conditionis referred to clinically as osteitis fibrosa.

Paget's disease (osteitis deformans) is a disorder currently thought tohave a viral etiology and is characterized by excessive bone resorptionat localized sites which flare and heal but which ultimately are chronicand progressive, and may lead to malignant transformation. The diseasetypically affects adults over the age of 25.

Although osteoporosis has been defined as an increase in the risk offracture due to decreased bone mass, none of the presently availabletreatments for skeletal disorders can substantially increase the bonedensity of adults. A strong perception exists among physicians thatdrugs are needed which could increase bone density in adults,particularly in the bones of the wrist, spinal column and hip that areat risk in osteopenia and osteoporosis.

Current strategies for the prevention of osteoporosis may offer somebenefit to individuals but cannot ensure resolution of the disease.These strategies include moderating physical activity, particularly inweight-bearing activities, with the onset of advanced age, includingadequate calcium in the diet, and avoiding consumption of productscontaining alcohol or tobacco. For patients presenting with clinicalosteopenia or osteoporosis, all current therapeutic drugs and strategiesare directed to reducing further loss of bone mass by inhibiting theprocess of bone absorption, a natural component of the bone remodelingprocess that occurs constitutively.

For example, estrogen is now being prescribed to retard bone loss. Thereis, however, some controversy over whether there is any long termbenefit to patients and whether there is any effect at all on patientsover 75 years old. Moreover, use of estrogen is believed to increase therisk of breast and endometrial cancer. High doses of dietary calciumwith or without vitamin D have also been suggested for postmenopausalwomen. However, ingestion of high doses of calcium can often haveunpleasant gastrointestinal side effects, and serum and urinary calciumlevels must be continuously monitored.

Other therapeutics which have been suggested include calcitonin,bisphosphonates, anabolic steroids and sodium fluoride. Suchtherapeutics however, have undesirable side effects, for example,calcitonin and steroids may cause nausea and provoke an immune reaction,bisphosphonates and sodium fluoride may inhibit repair of fractures,even though bone density increases modestly, which that may preventtheir usage.

The above disorders are examples of conditions that may lead to bonefractures, fissures or splintering of the bones in the individuals whosuffer from a given disorder. Current therapeutic methods areinsufficient to treat the disorders leaving a need for improvedtreatments of bone fractures when they occur in the individual. Thepresent invention provides improved compositions, products and methodsfor locally treating bone fractures, fissures, splintering and similarbreakages of the bone, or by strengthening decomposed bone tissue byincreasing the mechanism of mineralization of the bone. It isconceivable that the current invention also causes mineralization of thesurrounding connective tissue, such as collagen, cartilage, tendons,ligaments and other dense connective tissue and reticular fibers.

The Oral Cavity

With respect to tissue decomposition in the oral cavity, it is commonlyknown in the dental art that certain kinds of tooth decomposition anddecay that occurs over time in the mouth is initiated by acid etching ofthe tooth enamel with the source of the acid being a metaboliteresulting from bacterial and enzymatic action on food particles in theoral cavity. It is generally understood that plaque, a soft accumulationon the tooth surface consisting of an organized structure ofmicroorganisms, proteinaceous and carbohydrate substances, epithelialcells, and food debris, is a contributory factor in the development ofvarious pathological conditions of the teeth and soft tissue of the oralcavity. The saccharolytic organisms of the oral cavity which areassociated with the plaque, cause a demineralization or decalcificationof the tooth beneath the plaque matrix through metabolic activity whichresults in the accumulation and localized concentration of organicacids. The etching and demineralization of the enamel may continue untilthey cause the formation of dental caries and periodontal disease withinthe oral cavity.

Teeth are cycled through periods of mineral loss and repair also as aresult of pH fluctuations in the oral cavity. The overall loss or gainof mineral at a given tooth location determine whether the cariousprocess will regress, stabilize or advance to an irreversible state.Numerous interrelated patient factors affect the balance between theremineralization and demineralization portions of this cycle and includeoral hygiene, diet, and the quantity and quality of saliva. At the mostextreme point in this process, a restoration will be required to repairthe tooth.

Methods for the prevention and reduction of plaque and tooth decaywithin the oral cavity commonly involve the brushing of the teeth usingtoothpastes; mechanical removal of the plaque with dental floss;administration and rinsing of the oral cavity with mouthwashes,dentifrices, and antiseptics; remineralization and whitening of theteeth with fluoride agents, calcium agents and whitening agents, andvarious other applications to the oral cavity. Still missing in thefield is a delivery system for the remineralization of teeth that wouldaddress the challenges of demineralization facing the teeth continuallyin the oral cavity.

A tooth that has reached an advanced stage of decay often requiresinstallation of a dental restoration within the mouth. Half of alldental restorations fail within 10 years, and replacing them consumes60% of the average dentist's practice time. Current dental materials arechallenged by the harsh mechanical and chemical environment of the oralcavity with secondary decay being the major cause of failure.Development of stronger and longer-lasting biocompatible dentalrestorations by engineering novel dental materials or new resin systems,enhancing existing materials, and incorporating bioactive agents inmaterials to combat microbial destruction and to sustain the harshmechanical and chemical environment of the oral cavity continues to bedesired.

Despite numerous preventive oral health strategies, dental cariesremains a significant oral health problem. More than 50% of childrenaged 6-8 will have dental caries and over 80% of adolescents over age 17will have experienced the disease. Caries is also seen in adults both asa primary disease and as recurrent disease in already treated teeth.Advances in diagnosis and treatment have led to noninvasiveremineralizing techniques to treat caries. However mechanical removal ofdiseased hard tissue and restoration and replacement of enamel anddentin is still the most widely employed clinical strategy for treatingprimary caries, restoring function to the tooth and also blockingfurther decay. In addition, nearly 50% of newly placed restorations arereplacement of failed restorations. Clearly, restorative materials are akey component of treating this widespread disease.

The selection of a restorative material has significantly changed inrecent years. While dental amalgam is still considered a cost effectivematerial, there is a growing demand for tooth colored alternatives thatwill provide the same clinical longevity that is enjoyed by dentalamalgam. The use of composite resins has grown significantlyinternationally as a material of choice for replacing amalgam as arestorative material for posterior restorations. This demand ispartially consumer driven by preference for esthetic materials and theconcerns regarding the mercury content of amalgam. It is also driven bydentists recognizing the promise of resin-based bonded materials inpreserving and even supporting tooth structure. Numerous studies havesuggested that bonding the restoration to the remaining tooth structuredecreases fracture of multisurface permanent molar preparations.Unfortunately, posterior teeth restored with direct resin restorativematerials have a higher incidence of secondary caries. This has led toshorter clinical service and narrower clinical indications for compositeresin materials compared to amalgam.

The most frequently cited reason for restoration replacement isrecurrent decay around or adjacent to an existing restoration. It islikely that fracture at the margin due to polymerization shrinkage canlead to a clinical environment at the interface between a restorationand the tooth that collects dental plaque and thus promotes decay.Therefore, developing dental materials with anticaries capability is avery high priority for extending the longevity of restorations.

Tooth Remineralization

Although natural remineralization is always taking place in the oralcavity, the level of activity varies according to conditions in themouth as discussed. Incorporation of fluoride during theremineralization process has been a keystone for caries prevention. Theeffectiveness of fluoride release from various delivery platforms,including certain dental restorative materials has been widelydemonstrated. It is commonly accepted that caries prevention fromfluoride is derived from its incorporation as fluorapatite or fluorideenriched hydroxyapatite in the tooth mineral thereby decreasing thesolubility of tooth enamel. More recently, anticaries activity has beendemonstrated using the strategy of increasing solution calcium andphosphate concentrations to levels that exceed the ambient concentrationin oral fluids. In order for fluoride to be effective at remineralizingpreviously demineralized enamel, a sufficient amount of calcium andphosphate ions must be available. For every two (2) fluoride ions, ten(10) calcium ions and six (6) phosphate ions are required to form a cellof fluorapatite (Ca₁₀(PO₄)₆F₂). Thus the limiting factor for net enamelremineralization is the availability of calcium and fluoride in saliva.

The low solubility of calcium phosphates has limited their use inclinical delivery platforms, especially when in the presence of fluorideions. These insoluble phosphates can only produce available ions fordiffusion into the enamel in an acidic environment. They do noteffectively localize to the tooth surface and are difficult to apply inclinically usable forms. Because of their intrinsic solubility, solublecalcium and phosphate ions can only be used at very low concentrations.Thus they do not produce concentration gradients that drive diffusioninto the subsurface enamel of the tooth. The solubility challenge isexacerbated by the even lower solubility of calcium fluoride phosphates.

Several commercially available approaches exist using calcium andphosphate preparations that have been commercialized into various dentaldelivery models. These have been reportedly compounded to overcome thelimited bioavailability of calcium and phosphate ions for theremineralization process. The first technology uses caseinphosphopeptide (CCP) stabilized with amorphous calcium phosphate (ACP)(RECALDENT® CCP-ACP of Cadbury Enterprises Pte. Ltd.). It ishypothesized that the casein phosphopeptide can facilitate thestabilization of high concentrations of ionically available calcium andphosphate even in the presence of fluoride. This formulation binds topellicle and plaque and while the casein phosphopeptide prevents theformation of dental calculus, the ions are available to diffuse down theconcentration gradient to subsurface enamel lesions facilitatingremineralization. As compared to the CCP-ACP, in the composition of theinvention, biologically available ions are available due to the factthat the salts are already solvated in the microcapsule of theinvention. Amorphous calcium phosphate is not soluble in water orsaliva. Although the manufacturer claims release of bioavailable ionsfrom amorphous calcium phosphate, it is not as a result of thedissolution of the complex. A second technology (ENAMELON®) usesunstabilized amorphous calcium phosphate. Calcium ions and phosphateions are introduced as a dentifrice separately in a dual chamber deviceforming amorphous calcium phosphate in-situ. It is proposed thatformation of the amorphous complex promotes remineralization. A thirdapproach uses a so-called bioactive glass (NOVAMIN® of NovaMinTechnology Inc.) containing calcium sodium phosphosilicate. It isproposed that the glass releases calcium and phosphate ions that areavailable to promote remineralization. More recently dental compositeformulations have been compounded using zirconia-hybridized ACP that mayhave the potential for facilitating clinical remineralization.

While the Recaldent® and Enamelon® preparations have both in-situ andin-vivo evidence suggesting enhanced remineralization, these aretopically applied and do not specifically target the most at risklocation for recurrent caries at the tooth restoration interface. Whilethe bioactive glass and the zirconia-hybridized-ACP filler technologieshave potential, they are relatively inflexible in terms of the range offormulations in which they might be used due to the challenges ofdealing with brittle fillers and some of the limitations on controllingfiller particle size.

Another approach taken to decrease caries in the oral cavity is thelimiting of demineralization of enamel and bone by drinking waterfluoridation. It has been shown that the fluoride contained in drinkingwater incorporates to some extent into hydroxyapatite, the majorinorganic component of enamel and bone. Fluoridated hydroxyapatite isless susceptible to demineralization by acids and is thus seen to resistthe degradation forces of acidic plaque and pocket metabolites. Inaddition, fluoride ion concentration in saliva is increased throughconsumption of fluoridated drinking water. Saliva thus serves as anadditional fluoride ion reservoir and in combination with bufferingsalts naturally found in salivary fluid, fluoride ions are activelyexchanged on the enamel surface, further offsetting the effects ofdemineralizing acid metabolites.

Notwithstanding the established benefits of fluoride treatment of teeth,fluoride ion treatment can result in irregular spotting or blotching ofthe teeth depending on the individual, whether administered throughdrinking water or by topically applied fluoride treatment. This effectis known to be both concentration related and patient specific. Inaddition, the toxicology of fluoride is being studied as to its longterm effect on human health. Desired is a targeted approach offluoridation in the oral cavity.

Another approach to limiting the proliferation of microflora in the oralenvironment is through topical or systematic application ofbroad-spectrum antibacterial compounds. Reducing the number of oralmicroflora in the mouth results in a direct reduction or elimination ofplaque and pocket accumulation together with their damaging acidicmetabolite production. The major drawback to this particular approach isthat a wide variety of benign or beneficial strains of bacteria arefound in the oral environment which may be killed by the sameantibacterial compounds in the same manner as the harmful strains. Inaddition, treatment with antibacterial compounds may select for certainbacterial and fungi, which may then become resistant to theantibacterial compound administered and thus proliferate, unrestrainedby the symbiotic forces of a properly balanced microflora population.Thus the application or administration of broad-spectrum antibioticsalone is ill-advised for the treatment of caries and a more specific,targeted approach is desired.

Tooth Whitening

Cosmetic dental whitening or bleaching has become extremely desirable tothe general public. Many individuals desire a “bright” smile and whiteteeth, and consider dull and stained teeth cosmetically unattractive.Unfortunately, without preventive or remedial measures, stained teethare almost inevitable due to the absorbent nature of dental material.Everyday activities such as eating, chewing, or drinking certain foodsand beverages (in particular coffee, tea, and red wine) and smoking orother oral use of tobacco products cause undesirable staining ofsurfaces of teeth. Extrinsic staining of the acquired pellicle arises asa result of compounds such as tannins and polyphenolic compounds whichbecome trapped in and tightly bound to the proteinaceous layer on thesurfaces of teeth. This type of staining can usually be removed bymechanical methods of tooth cleaning. In contrast, intrinsic stainingoccurs when staining compounds penetrate the enamel and even the dentinor arise from sources within the tooth. The chromogens or color causingsubstances in these materials become part of the pellicle layer and canpermeate the enamel layer. Even with regular brushing and flossing,years of chromogen accumulation can impart noticeable toothdiscoloration. Intrinsic staining can also result from microbialactivity, including that associated with dental plaque. This type ofstaining is not amenable to mechanical methods of tooth cleaning andchemical methods are required.

Without specifically defining the mechanism of action of the presentinvention, the compositions, products and methods of the presentinvention enable the precipitation of salts onto the surfaces of theteeth in the oral cavity and make the salts available for adherence tothe tooth surface and remineralization of the teeth. The mineralizingsalts are deposited in the interstitial spaces of the teeth, making theteeth smoother, increasing the reflection of light from the surface ofthe teeth and thereby giving the teeth a brighter, more lustrousappearance and whiter visual effect.

Tooth whitening compositions generally fall into two categories: (1)gels, pastes, varnishes or liquids, including toothpastes that aremechanically agitated at the stained tooth surface in order to affecttooth stain removal through abrasive erosion of stained acquiredpellicle; and (2) gels, pastes, varnishes or liquids that accomplish thetooth whitening effect by a chemical process while in contact with thestained tooth surface for a specified period, after which theformulation is removed. In some cases, the mechanical process issupplemented by an auxiliary chemical process which may be oxidative orenzymatic. Initially, tooth whitening had been performed at thedentist's office. Less expensive at-home dental whitening kits havebecome available, such as whitening strips and whitening trays that comein either single compartment or dual compartment systems.

Both in-office and at-home tooth whitening typically involves theapplication of a peroxide containing composition to the surface of thetooth to achieve the desired whitening effect. The majority of mostin-office and at-home tooth whitening compositions act by oxidation.These compositions are applied directly by a patient in a toothbleaching tray, held in place in the mouth for contact times, sometimesfor periods of half an hour several times per day; or of greater than 60minutes per day, and sometimes as long as 8 to 12 hours. The slow rateof bleaching is, in large part, the consequence of formulations that aredeveloped to maintain stability of the oxidizing composition. Aqueoustooth whitening gels have proven desirable due to the hydrating effectson the structure of the tooth, reducing the likelihood of toothsensitivity.

The most commonly used oxidative compositions contain the hydrogenperoxide precursor carbamide peroxide which is mixed with an anhydrousor low water content, hygroscopic viscous carrier containing glycerineand/or propylene glycol and/or polyethylene glycol. When contacted bywater, carbamide peroxide dissociates into urea and hydrogen peroxide.The latter has become the tooth bleaching material of choice due to itsability to whiten teeth faster than higher concentrations of carbamideperoxide.

An alternative source of hydrogen peroxide is sodium percarbonate andthis has been used in a silicone polymer product that is painted ontothe teeth forming a durable film for overnight bleaching procedures. Theperoxide is slowly released for up to 4 hours.

Associated with the slow rate of bleaching in the hygroscopic carrier,the currently available tooth bleaching compositions cause toothsensitization in over 50% of patients. Tooth sensitivity is believed toresult from the movement of fluid through the dentinal tubes towardnerve endings in the tooth. This movement is enhanced by the carriersfor the carbamide peroxide. It has been determined that glycerine,propylene glycol and polyethylene glycol can each give rise to varyingamounts of tooth sensitivity following exposure of the teeth to heat,cold, overly sweet substances, and other causative agents.

Hydrogen peroxide tooth bleaching formulations have limitations inaddition to tooth sensitivity. Until recent years, stable aqueoushydrogen peroxide tooth bleaching gels have been virtually nonexistent.Hydrogen peroxide is a powerful oxidizing agent and an unstable compoundthat decomposes readily over time into water and oxygen. Certainchemical and physical influences in the oral cavity can accelerate therate of decomposition and need to be controlled for a stable toothwhitening gel to exist. Temperature, pH and errant metal ions all have aprofound effect on the decomposition of hydrogen peroxide, particularlyin an aqueous formula.

One advantage of the compositions of the invention is the decrease orelimination of tooth sensitivity of the patient. When used inconjunction with current tooth bleaching products, the microcapsules ofthe invention release salt ions that precipitate as salts in the oralcavity and mineralize the open dentin tubules of the teeth therebydecreasing tooth sensitivity to the oxidative tooth bleaching product.

Whitening systems on the market include two-part systems that requiremixing of the components upon administration and single partcompositions that are faster and easier to administer and generallypreferred for in-office bleaching by dentists. Two-part systems includeproducts such as dual barrel syringes, liquid hydrogen peroxide/powdersystems and whitening strips. Single component tooth bleachingcompositions prefer room temperature storage conditions in order toeliminate costly and inconvenient storage problems. The pH of an aqueoushydrogen peroxide tooth whitening composition also has great bearing onthe stability of the formulation. The two-part systems demonstratesuperior shelf life stability. Formulations that contain hydrogenperoxide solutions are strongly acidic and maintain their stability inacidic pH formulas. Stable aqueous hydrogen peroxide tooth whiteninggels can be formulated in the acid pH range. However, bleachingcompositions in the acidic pH range (pH 2.0-5.5) are prone to thedemineralization of dental enamel by the solubilizing of calcium ionsfrom the tooth surface. This reduction in surface enamel leads to toothsensitivity and discomfort for the patient. By incorporating thecompositions of the invention into tooth bleaching products or utilizingthem in conjunction with tooth bleaching products, the microcapsules ofthe invention can modify the pH level in the oral cavity to causeacceleration of the bleaching process.

Many of the available products are time-consuming and limited in theireffectiveness and subject the user to various physical discomforts. Moreimportantly, it has been shown that prolonged exposure of teeth towhitening compositions, as practiced at present, has a number of adverseeffects in addition to that of tooth sensitivity. Over time, any of theperoxides known in the art to achieve a desired tooth bleaching effectwill function as calcium chelating agents. Other examples of chelationagents often found in tooth whitening products include EDTA and itssalts, citric acid and its salts, gluconic acid and its salts, alkalimetal pyrophosphates and alkali metal polyphosphates. Solubilization ofcalcium from the enamel layer can occur at a pH less than 5.5 withassociated demineralization. The chelating agents will penetrate theintact enamel and dentin so as to reach the pulp chamber of a vitaltooth thereby risking damage to pulpal tissue. Other adverse effectsinclude dilution of the bleaching compositions with saliva in the oralcavity with resulting leaching from the dental tray and subsequentdigestion by the user.

It has been shown that the rate of whitening can be increased byincreasing the temperature of the hydrogen peroxide system, whereincrease of 10° C. can double the rate of reaction. Consequently, thereexist a number of procedures that utilize high intensity light to raisethe temperature of the hydrogen peroxide to accelerate the rate ofbleaching of the teeth. Other approaches to heating the hydrogenperoxide have been described such as the heating of dental instruments.Contemporary approaches and literature have focused on acceleratingperoxide bleaching with simultaneous illumination of the anterior teethwith various sources having a range of wavelengths and spectral power,for example, halogen curing lights, plasma arc lamps, lasers andlight-emitting diodes. Some products that are used in light activatedwhitening procedures contain ingredients that serve as photosensitizersthat claim to aid the energy transfer from the light to the peroxide geland are often colored materials, for example carotene and manganesesulfate. However, excessive heating can cause irreversible damage to thedental pulp. In addition, the literature for in vitro and clinicalstudies and actual results demonstrate that the actual effect of lighton tooth whitening is limited, conflicting and controversial.

There is thus a need for improved compositions, methods and productsthat overcome the limitations of the prior art. The challenge remains tocreate a tooth whitening and remineralization technology platform forincorporating stable and effective tissue remineralization ions that canbe incorporated into a myriad of dental materials and variety ofproducts. Such a delivery platform would facilitate the formulation ofdental products capable of remineralization of the teeth. Thecompositions, products and methods of the current inventions asdescribed herein satisfy these and other needs. The ultimate impact is areduction in recurrent caries, the most prevalent reason for restorationreplacement; whitening of the teeth; and resulting improvement inoverall strength and health of the teeth in the oral cavity.

There exists a broad need for improved compositions and methods usefulfor therapeutic agent delivery. In particular, there is a need for animproved microcapsule-based technology for delivering therapeutic agentsto diverse tissue types in a stable and time-controlled manner.

SUMMARY OF THE INVENTION

In accordance with the description herein and desire to provide improvedtherapeutic products, the present invention provides compositions andmethods that deliver a buffered therapeutic agent in a controlledfashion. Also presented are methods for using such compositions in thetreatment and prevention of a wide variety of conditions resulting frommicroorganisms. The invention also provides compositions and productsfor antimicrobial coatings. More particularly, the present inventionprovides a composition comprising buffered solutions of therapeuticagents encapsulated in a semipermeable polymer shell that allows therelease of therapeutic agents to be delivered to a patient. Themicrocapsules can be incorporated into a variety of products asdiscussed herein, and can be prepared by any generally knownmicroencapsulation method, but preferably by surfactant-free inverseemulsion. More particularly, the invention includes a compositioncontaining polymer microencapsulated solutions of buffered therapeuticagents. Further, the rate of release of the therapeutic agent from themicrocapsules can be designed in a single type of microcapsule and in aproduct containing a number of different types of microcapsules. Thisresults in controlled time release of the agent and thereby atherapeutic effect over a prolonged period of time.

A preferred embodiment is directed towards a topical microcapsulecomposition comprising: a plurality of microcapsules and a carrier, eachof said microcapsules comprising at least one polymer substantiallydisposed as a semipermeable shell around a buffered solution, saidmicrocapsule composition comprising at least a first subset ofmicrocapsules, wherein a microcapsule within said first subset ofmicrocapsules comprises a therapeutic agent within said microcapsule,for topical use on epithelial tissue; wherein the therapeutic agentpermeates the shell.

A further preferred embodiment is towards the microcapsule composition,wherein the buffered solution is an aqueous solution.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the therapeutic agent is selected from the groupconsisting of antibacterial agents, antifungal agents, benzoyl peroxide,coal tar, corticosteroids, retinoids, salicyclic acid, antiviral agents,immunosuppressants, and biologic agents for treating psoriasis.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the composition is a moisturizer, cream, lotion, foam,gel, shampoo, hair conditioner, hair gel, or a nail polish.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the molecular weight of the polymer is from about 1,000to about 50,000.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the polymer is cross-linked.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the microcapsule has a diameter of from about 1 micronto about 3 mm.

A further preferred embodiment is towards the microcapsule composition,further comprising a second subset of microcapsules, wherein the secondsubset of microcapsules comprises within each of said second subset ofmicrocapsules, a second therapeutic agent in a buffered solution andencapsuled within a semipermeable shell, wherein the second therapeuticagent permeates the semipermeable shell, and wherein the composition issuitable for delivery to a mammal. In a further preferred embodiment,wherein the first subset of microcapsules has a therapeutic agentrelease profile that differs from the therapeutic agent release profileof the second subset of microcapsules.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the microcapsule is made of a polymeric system thatundergoes a sol-gel transition once administered.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the microcapsule is biodegradeable.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the polymer is an amphiphilic polyurethane polymer.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the therapeutic agent is a natural antimicrobial agentselected from the group consisting of: beta lactam antiobiotics such aspenicillins or cephalosporins, protein synthesis inhibitors such asaminoglycosides, macrolides, ketolides, tetracyclines, chloramphenicol,and polypeptides; penicillins including: penicillin G, procainepenicillin, benzathine penicillin, and penicillin V; cephalosporinsincluding: cefacetrile, cefadroxil, cephalexin, cefaloglycin,cefalonium, cefaloridine, cefalotin, cefapirin, cefatrizine, cefazaflur,cefazedone, cefazolin, cefradine, cefroxadine, ceftezole, cefaclor,cefonicid, cefprozil, cefuroxime, cefuzonam, cefmetazole, cefotetan andcefoxitin; aminoglycosides including: amikacin, arbekacin, gentamicin,kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin,streptomycin, tobramycin, and apramycin; Macrolides including:azithromycin, clarithromycin, dirithromycin, erythromycin,roxithromycin, and telithromycin; Ketolides including: telithromycin,cethromycin, solithromycin, spiramycin, ansamycin, oleandomycin,carbomycin and tylosin; naturally occurring tetracyclines including:tetracycline, chlortetracycline, oxytetracycline, and demeclocycline;semisynthetic tetracyclines including: doxycycline, lymecycline,meclocycline, methacycline, minocycline and rolitetracycline;polypeptides including: actinomycin, bacitracin, colistin, and polymyxinB; synthetic antimicrobial agents including: sulphonamides,cotrimoxazole, quinolones, antivirals, antifungals, anticancer drugs,antimalarials, antituberculosis drugs, antileprotics and antiprotozoals;sulphonamide antibacterials including: sulfamethoxazole, sulfisomidine,sulfacetamide, sulfadoxine, dichlorphenamide, and dorzolamide;Sulphonamide diuretics including: acetazolamide, bumetanide,chlorthalidone, clopamide, furosemide, hydrochlorothiazide, indapamide,mefruside, metolazone, and xipamide; sulphonamide anticonvulsantsincluding: acetazolamide, ethoxzolamide, sultiame and zonisamide; andother sulfonamide therapeutic agents including: celecoxib, darunavir,probenecid, sulfasalazine, and sumatriptan.

A further preferred embodiment is towards the microcapsule composition,wherein wherein the therapeutic agent is an antifungal agent selectedfrom: amphotericin B, candicidin, filipin, hamycin, natamycin, nystatin,rimocidin; and imidazole, triazole, and thiazole types including:bifonazole, butoconazole, clotrimazole, econazole, fenticonazole,isoconazole, ketoconazole, miconazole, omoconazole, oxiconazole,sertaconazole, sulconazole, tioconazole, albaconazole, fluconazole,isavuconazole, itraconazole, posaconazole, ravuconazole, terconazole,voriconazole and abafungin; and ecninocandins including anidulafungin,caspofungin, and micafungin.

A further embodiment is directed towards a method of forming a topicalmicrocapsule composition comprising a plurality of microcapsules: saidplurality of microcapsules being formed by combining a polymer and atleast one buffered aqueous solution comprising a therapeutic agent, bycontacting: (a) said buffered solution containing the therapeutic agent,(b) an oil phase, (c) said polymer, and (d) an emulsifying agent,wherein the polymer substantially forms a semipermeable shell around thebuffered aqueous solution; and adding said plurality of microcapsules toa carrier.

A further preferred embodiment is directed towards the method above,wherein said plurality of microcapsules comprises a first and a secondplurality of microcapsules, wherein a first plurality of microcapsulesis formed using a first therapeutic agent, and a second plurality ofmicrocapsules is formed using a second therapeutic agent.

A further preferred embodiment is directed towards the method above,wherein the first and second plurality of microcapsules have a differentrelease profile.

A further preferred embodiment is directed towards the method above,further comprising a diol, an isocyanate or both, wherein said diol,isocyanate, or both increases the molecular weight of an isocyanatefunctionalized polyurethane shell.

A further preferred embodiment is directed towards the method above,wherein the oil phase is methyl benzoate.

A further preferred embodiment is directed towards the method above,wherein the emulsifying agent is a polyglyceryl-3-polyricinoleate.

A further preferred embodiment is directed towards the method above,wherein said therapeutic agent is selected from the group consisting ofantibacterial agents, antifungal agents, benzoyl peroxide, coal tar,corticosteroids, retinoids, salicyclic acid, antiviral agents,immunosuppressants, and biologic agents for treating psoriasis.

A further preferred embodiment is directed towards the method above,wherein the carrier is in the form of a moisturizer, cream, lotion,foam, gel, shampoo, hair conditioner, hair gel, or a nail polish.

A further preferred embodiment is directed towards the method above,wherein the therapeutic agent is a biologically active additive or apeptide-based or peptide analog-based antimicrobial agent.

A further preferred embodiment is directed towards the method above,wherein the therapeutic agent is a natural or synthetic oil: flavoringoils, flavoring aldehydes, esters, alcohols, similar materials, andcombinations thereof, vanillin, sage, marjoram, parsley oil, spearmintoil, cinnamon oil, oil of wintergreen (methyl salicylate), peppermintoil, clove oil, bay oil, anise oil, eucalyptus oil, citrus oils, fruitoils and essences including those derived from lemon, orange, lime,grapefruit, apricot, banana, grape, apple, strawberry, cherry,pineapple, bean- and nut-derived flavors such as coffee, cocoa, cola,peanut, and almond.

A further preferred embodiment is directed towards the method above,wherein the therapeutic agent is selected from the group consist of:menthol, menthyl acetate, menthyl lactate, camphor, eucalyptus oil,eucalyptol, anethole, eugenol, cassia, oxanone, a-irisone, propenylguaiethol, thymol, linalool, benzaldehyde, cinnamaldehyde,N-ethyl-p-menthan-3-carboxamine, N,2,3trimethyl-2-isopropylbutanamide,3-1-menthoxypropane-1,2-diol, cinnamaldehyde glycerol acetal (CGA),methone glycerol acetal (MGA), and mixtures thereof.

A further preferred embodiment is directed towards the method above,wherein the therapeutic agent is selected from the group consist of:butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), vitaminA, carotenoids, vitamin E, flavonoids, polyphenols, ascorbic acid,herbal antioxidants, chlorophyll, melatonin, and mixtures thereof.

A further preferred embodiment is directed towards the method above,wherein the therapeutic agent is selected from the group consisting of:Halogenated hydrocarbons selected from: salicylanilides, carbanilides,bisphenols, diphenyl ethers, anilides of thiophene carboxylic acids andchlorhexidines; Quaternary ammonium compounds selected from: alkylammonium, pyridinum, and isoquinolinium salts, and Sulfur activecompounds selected from: thiuram sulfides and dithiocarbamates, andcombinations thereof.

A further preferred embodiment is directed towards the method above,wherein the carrier is in the form of a moisturizer, cream, lotion,foam, gel, shampoo, hair conditioner, hair gel, or a nail polish.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an epifluorescent microscopic image of a microcapsulecontaining FITC-labeled lysozyme in PBS. The imaged microcapsule isapproximately 7 micrometers in size.

FIG. 2 is a confocal fluorescent imaging scan of a microcapsulecontaining FITC-labeled lysozyme in PBS. The imaged microcapsule isapproximately 7 micrometers in size. Without the use of a buffersolution, the lysozyme denatures and fluorescence is not observed.

DETAILED DESCRIPTION OF THE INVENTION

This invention provides buffered, as well as nonbuffered, microcapsulecompositions for delivering therapeutic agents to a mammal, preferably ahuman. The nonlimiting description below sets forth various embodimentsof the subject compositions, and methods of making and using same.

Production of Microcapsules

Microencapsulation

In a preferred embodiment, the composition of the invention is formed bycombining a buffered solution, therapeutic agent, oil-solubleemulsifying agent and at least one type of polymer which, when combinedand upon mixing or agitation, form the microcapsules of the invention.As used herein, the term “microcapsules” includes tiny particles ordroplets surrounded by a coating to give small capsules having usefulproperties. Microcapsules are sometimes referred to as microspheres,though a microcapsule of the invention need not be spherical in shape.The material inside the microcapsule shall be referred to herein by thesynonymous terms “core” and “internal phase,” and the materialsurrounding the core is referred to herein by the synonymous terms“shell,” “wall,” “coating,” “membrane” and “exterior phase.” The shellneed not be completely or uniformly placed around the core of themicrocapsule, as long as substantially all of the core is surrounded bya polymer shell, as will be described further herein.

Preferably, the microcapsules of the invention have a diameter range ofbetween 100 nanometers and 3 millimeters. More preferably, the size ofthe microcapsules is between 1 micron and 1 mm. In general, thepreferable size of the microcapsule will be governed by the desired enduse application. One parameter used to control the size of themicrocapsules is by the amount and force of mixing or agitation of theemulsion. Other parameters to control the size of the microcapsules andcomponents of the microcapsules will be discussed further below. Thesize of the instant microcapsules can be optimized so that a sufficientnumber of microcapsules are available to affect a therapeutic response.

Methods for constructing microcapsules may be physical or chemical.Physical methods include pan coating, air-suspension coating,centrifugal extrusion, vibrational nozzle and spray-drying. Chemicalmethods include polymerization such as interfacial polymerization,in-situ polymerization and matrix polymerization. In interfacialpolymerization, at least two monomers are dissolved separately inimmiscible liquids. Upon interface between the liquids, rapid reactionoccurs, creating a thin shell or wall of the microcapsule. In-situpolymerization is the direct polymerization of a single monomer carriedout on the particle's surface. In matrix polymerization, a core materialis embedded during formation of the microcapsule. Microcapsules mightalso be constructed by using sol-gel techniques, by aqueous or organicsolution precipitation synthesis methods, complex coacervation, and byother methods known in the art.

A preferred method of preparing the instant microcapsules is a synthesisto generate microcapsules containing buffer solutions of therapeuticagents, and in particular, antimicrobial agents. In order to encapsulatebuffered solutions of biologically active additives in a microcapsule, asurfactant-free inverse emulsion of water in oil is preferably used. Anycontinuous oil phase can be used for the process of the invention. Inone embodiment, hydrophobic oils are used as the continuous oil phasewithin the process with an emulsifying agent that serves to stericallystabilize the dispersed phase. One preferred oil phase of the inventionis methyl benzoate. FIG. 1 is an epifluorescent microscopic image of amicrocapsule containing FITC-labeled lysozyme in PBS. The imagedmicrocapsule is approximately 7 micrometers in size. FIG. 2 is aconfocal fluorescent imaging scan of a microcapsule containingFITC-labeled lysozyme in PBS. The imaged microcapsule is approximately 7micrometers in size.

A standard emulsion uses a surfactant to stabilize a dispersed droplet,whereas the present preferred method uses an emulsifying agent in acontinuous oil phase to sterically stabilize the dispersed waterdroplets in order to allow an interfacial polymerization to occur. Thiscauses an effective synthesis of polymer shells around the bufferedtherapeutic agent solution in the dispersed phase. The amphiphiliccharacter of surfactants causes interference with the polymerizationthat needs to occur at the interface of the dispersed phase and thecontinuous phase that is necessary to generate a capsule. A surfactantalso presents a problem in its affinity for ions. The polar hydrophilichead group could be attracted to certain types of the therapeutic agentsor the ions contained in the capsule for buffering the biologicallyactive additive. The presence of a surfactant decreases the percentageof, for example, ionic therapeutic agents that are truly bioavailable,effectively behaving as a chelation agent inactivating the release ofthe therapeutic agent from the capsule if the therapeutic agent is ionicin nature. Consequently, surfactant-free inverse emulsion interfacialpolymerization is preferred as the method for forming the instantmicrocapsules of the invention.

Emulsifying Agents

The emulsifying agents preferred in the microcapsules of the inventionare different from surfactants in that the emulsifying agentsexclusively partition into the oil phase and are not surface active.Inherent in the concept of using surfactant-free inverse emulsions isthat water droplets can be disrupted into small droplets, the size andsize distribution of which are dependent on the form and amount of inputenergy, and the droplets formed survive transiently due to a rathersluggish growth rate. Although surfactant-free emulsions have beenfrequently applied in solvent extraction, emulsion polymerization, andfood production such as oil-and-vinegar dressing production, very littleattention has been paid to its fundamental properties for use inmicroencapsulating buffered aqueous therapeutic agent solution systems.The emulsifying agent sterically stabilizes droplets without interferingwith interfacial polymerization.

Polymers

The microcapsules of the invention contain a shell comprised of at leastone polymer, preferably with the shell being semipermeable to particulartherapeutic agents, whether in buffered solutions. As used herein, theterms “polymer” and “polymers” are intended to connote precursor polymermolecules having a preferable size in the range of 1,000 to 50,000g/mole; more preferably from 1,500 to 20,000 g/mole; and more preferablyfrom 1,500 to 8,000 g/mole. Larger polymers can be used, as well assmaller oligomers or prepolymers, but the molecular weight of thepolymer is controlled for practical uses in the desired productapplications. Conceivably, monomers can be used as well in the method ofthe invention. A number of polymers can be combined into onemicrocapsule in order to produce an end use product having particularlydesired release characteristics of the core components.

In one embodiment of the invention, the microcapsule shell is designedwith limited or substantially no permeability depending on its desiredapplication. The impermeable shell is formed during synthesis byselecting particular polymers known to be impermeable to the particulartherapeutic agents in the desired end use application. Suchmicrocapsules may, for example, be synthesized for “burst” applicationas discussed herein.

Many classes of polymers can be used in the scope of the invention andthe choice depends on the specific desired properties. Examples include,but are not limited to, acrylic polymers, alkyl resins, aminoplasts,coumarone-indene resins, epoxy resins, fluoropolymers, phenolic resins,polyacetals, polyacetylenes, polyacrylics, polyalkylenes,polyalkenylenes, polyalkynylenes, polyamic acids, polyamides,polyamines, polyanhydrides, polyarylenealkenylenes,polyarylenealkylenes, polyarylenes, polyazomethines, polybenzimidazoles,polybenzothiazoles, polybenzoxazinones, polybenzoxazoles, polybenzyls,polycarbodiimides, polycarbonates, polycarboranes, polycarbosilanes,polycyanurates, polydienes, polyester-polyurethanes, polyesters,polyetheretherketones, polyether-polyurethanes, polyethers,polyhydrazides, polyimidazoles, polyimides, polyimines,polyisocyanurates, polyketones, polyolefins, polyoxadiazoles,polyoxides, polyoxyalkylenes, polyoxyarylenes, polyoxymethylenes,polyoxyphenylenes, polyphenyls, polyphosphazenes, polypyrroles,polypyrrones, polyquinolines, polyquinoxalines, plysilanes,polysilazanes, polysiloxanes, polysilsesquioxanes, polysulfides,polysulfonam ides, polysulfones, polythiazoles, polythioalkylenes,polythioarylenes, polythioethers, polythiomethylenes,polythiophenylenes, polyureas, polyurethanes, polyvinyl acetals,polyvinyl butyrals, and polyvinyl formals. One skilled in the art willfurther appreciate that the selection of the specific type of polymerwill affect the composition and permeability characteristics of thesubject microcapsules.

Buffers

Buffers have a vast significance in all areas of science. A bufferedsolution resists changes in pH when acids or bases are added or whendilution occurs. A buffer is a mixture of an acid and its conjugatebase. There must be comparable amounts of the conjugate acid and base,within a factor of 10, to exert significant buffering. Buffers allow forthe proper functioning of any biological system and its subcellularcomponents such as proteins and peptide-based molecules. Nearly allbiological systems depend on pH. For example, a buffer solutionmaintains the correct pH for enzymes in many organisms to work.Typically, enzymes function only under very precise conditions. If thepH moves outside of a narrow range, the enzymes slow or stop functioningand can denature. pH directly affects the rate of enzyme-catalyzedreactions.

In some cases, the buffering of a therapeutic agent solution is requiredand in other cases the buffering of a therapeutic agent solution ispreferred to enhance shelf life and stability of a product. In somecases, the buffering of a therapeutic agent solution is preferred togain a more substantive effect of the agent. The rate of therapeuticagent release is a critical factor in patient care. This inventionprovides methods and compositions targeted at the microencapsulation ofa buffered therapeutic solution capable, for example, of a sustained,prolonged release of the therapeutic agent, a burst release of thetherapeutic agent, or a selective variable rate of release of thetherapeutic agent (e.g., a quick burst followed by sustained release, orsustained release followed by a quick burst).

Many classes of buffer solutions can be used in the invention, and thechoice of buffer depends on the specific pH desired. Examples include,but are not limited to, the following: Phosphate buffered saline whichis a solution containing sodium chloride, sodium phosphate, and, in someformulations, potassium chloride and potassium phosphate;3-([tris(hydroxymethyl)methyl]amino) propane-sulfonic acid (TAPS);N,N-bis(2-hydroxyethyl)glycine (Bicine);tris(hydroxyl-methyl)methylamine (Tris);N-tris(hydroxymethyl)methylglycine (Tricine);3-[N-Tris-(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid(TAPSO); 4-2-hydroxy-ethyl-1-piperazineethanesulfonic acid (HEPES);2-([tris(hydroxymethyl)methyl]amino) ethanesulfonic acid (TES);3-(N-morpholino)propanesulfonic acid (MOPS);piperazine-N,N′-bis(2-ethanesulfonic acid) (PIPES); dimethylarsinic acid(Cacodylate); saline sodium citrate (SSC);2-(N-morpholino)ethanesulfonic acid (MES); Phosphoric acid; citric acid;piperazine-N,N′-bis(3-propanesulfonic acid (PIPPS);piperazine-N,N′-bis(3-butanesulfonic acid) (PIPBS);N,N′-Diethylethylenediamine-N,N′-bis(3-propanesulfonic acid) (DESPEN);N,N′-diethylpiperazine dihydrochloride (DEPP.2HCl);N,N,N′,N′-tetraethyl-ethylenediamine dihydrochloride (TEEN.2HCl);N-2-Acetamidoiminodiacetic acid (ADA);1,3-Bis[tris(hydroxymethyl)methylamino]propane hydrochloride (BIS-TRISpropane.HCl); N-2-acetamido-2-aminoethanesulfonic acid (ACES);3-(N-Morpholino)-2-hydroxypropanesulfonic acid (MOPOS); imidazolehydrochloride; 3-(N-morpholino)butanesulfonic acid (MOBS);4-2-hydroxyethyl-1-piperazinepropane-sulfonic acid (HEPPS);N-tris(hydroxymethyl)methylglycine (TRICINE); glycine amidehydrochloride; Tris(hydroxymethyl)aminomethane hydrochloride (TRIShydrochloride); glycylglycine; Boric acid; cyclohexylaminoethanesulfonicacid (CHES); 3-(Cyclohexylamino)propane sulfonic acid (CAPS);N,N,N′,N′-tetraethylmethylene-diamine dihydrochloride (TEMN.2HCl); HCland sodium citrate; citric acid and sodium citrate; acetic acid andsodium acetate; K₂HPO₄ and KH₂PO₄; Na₂HPO₄ and NaH₂PO₄;N-cyclohexyl-2-aminoethanesulfonic acid; sodium borate; and sodiumhydroxide.

It will be known to those skilled in the art that the buffer pH dependson ionic strength and temperature. Compositions can be adjustedaccordingly during the synthesis, storage and use of themicroencapsulated buffer solution.

Surfactant-Free Inverse Emulsion Polymerization

In a preferred embodiment of surfactant-free inverse emulsionpolymerization, a very low molecular weight polyurethane is premixedinto the continuous oil phase. Preferably, the molecular weight of thepolyurethane is 1,500 to 20,000 g/mole, and more preferably from 1,500to 8,000 g/mole. Due to the amphiphilic nature of the low molecularweight polyurethane, the polyurethane spends the majority of the time atthe interface of the dispersed and continuous phases.

In this embodiment, diol is added to the system to increase themolecular weight of an isocyanate functionalized polyurethane shell. Apreferred diol is ethylene glycol. The diol ultimately leads to ethyleneoxide linker units in the microcapsule chemical structure. It has beenshown in industrial applications that ethylene oxide does not inhibitthe flow of ions between electrodes. This approach is useful forunderstanding the structure-property relationship of the urethane onmicrocapsule permeability due to the ease with which the chemicalstructure can be varied in the synthesis of the microcapsules by simplychanging the identity of the diol used in the polyurethane wall. In thisembodiment, the ion permeability of the microcapsule shell is based onthe chemical composition of the diols that act as spacer monomers. Thefollowing scheme represents the reaction used to synthesize themicrocapsule shell of this embodiment.

The length of the ethylene oxide spacer in the microcapsule wall may bevaried in order to control ion permeability of the membrane shell.Preferred embodiments include microcapsules using ethylene glycol (n=1)and from 1,4-butanediol (n=2). Preferred embodiments includemicrocapsules from diols where n=3 (1,6-hexanediol) or 4(1,8-octanediol). Preferred embodiments include microcapsules frompolymeric diols such as polyethylene glycol (also known as polyethyleneoxide or polyoxyethylene). Preferred embodiments include polyol monomerscapable of forming a crosslinked polymer microcapsule wall. Many classesof polyols can be used in the scope of the invention and the choicedepends on the specific desired properties. Examples include, but arenot limited to, pentaerythritol and glycerol.

In one embodiment, the microcapsule is biodegradable. The polyurethaneprepolymer could, for example, have a block of a polyester added to itto enhance biodegradation. Polylactic acid or polylactide could beincorporated into the microcapsule chemical structure to controlbiodegradation of the microcapsule.

A second characteristic influencing the permeability of the microcapsulemembrane is the shell or wall thickness, which may be varied by varyingthe ratio of the mass of material used to synthesize the shell to thevolume of the dispersed buffered therapeutic agent solution. At aconstant stir rate, adding more material relative to the bufferedaqueous therapeutic agent phase will lead to formation of thickermicrocapsule walls. In a preferred embodiment, the invention comprises aratio of 1 gram of polyurethane to from 15 to 40 mL of aqueous buffersolution.

Embodiments of the materials of the invention can be formulated suchthat only one type of buffered therapeutic agent is contained within thecore of the microcapsule, or alternatively, a plurality of differenttypes of buffered therapeutic agents, and can be incorporated into onemicrocapsule.

In other embodiments, a plurality of microcapsules containing one typeof buffered additive can be combined in a product with microcapsulescontaining other additives.

Loading of Microcapsules

The microcapsules of the invention contain semipermeable polymer shellswherein the permeability functions to release a therapeutic agent bothout of the microcapsules and into the microcapsules from the surroundingenvironment as a result of concentration gradients. Thus, embodimentsare contemplated where already formed microcapsules having none or lessthan the maximum possible amount of the therapeutic agent dissolved inthe buffered solutions in the core can be charged with additionaltherapeutic agent solution, herein referred to as “loading.” Loadingalso includes “recharging” of microcapsules with buffered solutionwithout therapeutic agents in the presence of the target bufferedtherapeutic agent and appropriate concentration gradients. The newtherapeutic agent can be introduced into the core of partially loadedmicrocapsules or reintroduced into the core of empty microcapsules byimmersing the microcapsules into buffered solutions of highly chargedtherapeutic agents where the concentration of the therapeutic agent inthe buffered solution is higher than the concentration of thetherapeutic agent in the buffer solution within the core of themicrocapsules. The recharge rate of the microcapsules can depend on butis not limited to the following variables including the concentrationgradients of the therapeutic agents, the temperature and the releaseprofile of the particular polymers of the product.

The method of loading a therapeutic agent into a microcapsule thatcontains only buffer solution is preferred when any heat above normalbody temperature is needed in the formation of the microcapsule. Thismethod avoids the use of heat that could cause a peptide-basedtherapeutic agent to denature.

Therapeutic Agents

As used herein, the term “therapeutic agent” means an agent (e.g., anatom, an ion, a salt, a molecule [such as an inorganic molecule, anorganic molecule or a biomolecule (e.g., a peptide, a peptide analog ora protein)], a solid or a liquid) that brings about a beneficial effectto a mammal or any tissue or other subpart thereof.

Many classes of therapeutic agents can be used in the invention. Indeed,these therapeutic agents are envisioned for use in treating a wide arrayof conditions, such as demineralization of bone; weakness of bone, skin,hair and nails; drying, staining and cracking of skin; tooth sensitivityand discoloration; skin dehydration; and infections of all types.Examples of therapeutic agents include, but are not limited to, thevarious agents set forth below.

Antimicrobial Agents

Antimicrobial agents are substances that kill or inhibit the growth ofmicroorganisms such as bacteria, fungi or protozoa. Antimicrobial agentscan be in the form of a drug that is either microbiocidal ormicrobiostatic. Antimicrobial agents can be used outside the human bodyor on nonliving objects. Antimicrobial agents that are drugs can comefrom either natural sources or can be synthetically prepared.

Antimicrobial agents can be used in many different products, such asmedical devices, wound dressings, fabrics, and numerous consumerproducts, both to protect the product from antimicrobial growth or forpublic health purposes. Natural or synthetic antimicrobial materialshave the potential to be included into other materials or used directlyon product surfaces or contained within a surface coating.

Natural antimicrobial agents include, but are not limited to, betalactam antiobiotics such as penicillins or cephalosporins, proteinsynthesis inhibitors such as aminoglycosides, macrolides, ketolides,tetracyclines, chloramphenicol, and polypeptides. Penicillins include,but are not limited to, penicillin G, procaine penicillin, benzathinepenicillin, and penicillin V. Cephalosporins include, but are notlimited to, cefacetrile, cefadroxil, cephalexin, cefaloglycin,cefalonium, cefaloridine, cefalotin, cefapirin, cefatrizine, cefazaflur,cefazedone, cefazolin, cefradine, cefroxadine, ceftezole, cefaclor,cefonicid, cefprozil, cefuroxime, cefuzonam, cefmetazole, cefotetan andcefoxitin. Aminoglycosides include, but are not limited to, amikacin,arbekacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin,rhodostreptomycin, streptomycin, tobramycin, and apramycin. Macrolidesinclude, but are not limited to, azithromycin, clarithromycin,dirithromycin, erythromycin, roxithromycin, and telithromycin. Ketolidesinclude, but are not limited to, telithromycin, cethromycin,solithromycin, spiramycin, ansamycin, oleandomycin, carbomycin andtylosin. Naturally occurring tetracyclines include, but are not limitedto, tetracycline, chlortetracycline, oxytetracycline, anddemeclocycline. Semisynthetic tetracyclines include, but are not limitedto, doxycycline, lymecycline, meclocycline, methacycline, minocyclineand rolitetracycline. Polypeptides include, but are not limited to,actinomycin, bacitracin, colistin, and polymyxin B. Syntheticantimicrobial agents include, but are not limited to, sulphonamides,cotrimoxazole, quinolones, antivirals, antifungals, anticancer drugs,antimalarials, antituberculosis drugs, antileprotics and antiprotozoals.Sulphonamide antibacterials may include, but are not limited to,sulfamethoxazole, sulfisomidine, sulfacetamide, sulfadoxine,dichlorphenamide, and dorzolamide. Sulphonamide diuretics include, butare not limited to, acetazolamide, bumetanide, chlorthalidone,clopamide, furosemide, hydrochlorothiazide, indapamide, mefruside,metolazone, and xipamide. Sulphonamide anticonvulsants include, but arenot limited to, acetazolamide, ethoxzolamide, sultiame and zonisamide.Other sulfonamide therapeutic agents include, without limitation,celecoxib, darunavir, probenecid, sulfasalazine, and sumatriptan.

Antifungal Agents

Common fungal infections include athlete's foot, ringworm, andcandidiasis. Fungi can also cause systemic infections like cryptococcalmeningitis. Antifungals work by exploiting differences between mammalianand fungal cells to kill off the fungal organism without dangerouseffects on the host. Unlike bacteria, both fungi and humans areeukaryotes.

Antifungal agents can be of the polyene type which includes, but is notlimited to, amphotericin B, candicidin, filipin, hamycin, natamycin,nystatin and rimocidin. Antifungal agents can be of the imidazole,triazole, and thiazole types which include, but are not limited to,bifonazole, butoconazole, clotrimazole, econazole, fenticonazole,isoconazole, ketoconazole, miconazole, omoconazole, oxiconazole,sertaconazole, sulconazole, tioconazole, albaconazole, fluconazole,isavuconazole, itraconazole, posaconazole, ravuconazole, terconazole,voriconazole and abafungin.

Antifungal agents can also be echinocandins which can include, but arenot limited to, anidulafungin, caspofungin and micafungin.

Antibacterial Agents

Host defense proteins and peptide-based antibiotic drugs can selectivelytarget and puncture the bacterial cell membrane. Host defense proteinswhich are part of the innate immune system in the body represent a firstline of defense against bacterial attack and primarily exist in therespiratory tract, urogenital tract, gastrointestinal track andepidermal tissues under the skin, all entry points for microbialpathogens in the body. These proteins kill bacteria by targetingbacterial membranes thereby generating an instability of the cellularcontents and membrane, which results in the death of the bacteria.Examples include the treatment of bacterial skin infections caused byStaphylococcus aureus, or the treatment of other blood streaminfections, lung infections and oral mucositis.

An approach to limiting the proliferation of microflora in the oralenvironment is through topical or systematic application ofbroad-spectrum antibacterial compounds. The reduction of the number oforal microflora in the mouth results in a direct reduction orelimination of plaque and pocket accumulation together with theirdamaging acidic metabolite production. The major drawback of thisparticular approach is that a wide variety of benign or beneficialstrains of bacteria are found in the oral environment which may bekilled by the same antibacterial compounds in the same manner as theharmful strains. In addition, treatment with antibacterial compounds mayselect for certain bacterial and fungi, which may then become resistantto the antibacterial compound administered and thus proliferate,unrestrained by the symbiotic forces of a properly balanced microflorapopulation. Thus, the application or administration of broad-spectrumantibiotics alone is ill-advised for the treatment of caries and a morespecific, targeted approach is desired.

The appearance of strains of drug-resistant bacteria and the increasednumber of drug-resistant infections has necessitated new approaches totreat these infections. There are many antibiotics in development fortreating resistant bacterial infections. Some of the emerging drugs arepeptide-based drugs and peptide analog-drugs. An effective method ofdelivery in an aqueous solution would be improved by storage in a buffersolution in order to improve shelf life and stability, along withleading to a more substantive effect of the therapeutic agent.

Additional antibacterial agents include, without limitation, copper (II)compounds such as copper (II) chloride, fluoride, sulfate and hydroxide,zinc ion sources such as zinc acetate, zinc citrate, zinc gluconate,zinc glycinate, zinc oxide, zinc sulfate and sodium zinc citrate,phthalic acid and salts thereof such as magnesium monopotassiumphthalate, hexetidine, octenidine, sanguinarine, benzalkonium chloride,domiphen bromide, alkylpyridinium chlorides such as cetylpyridiniumchloride (CPC) (including combinations of CPC with zinc and/or enzymes),tetradecylpyridinium chloride and N-tetradecyl-4-ethylpyridiniumchloride, iodine, halogenated carbanilides, halogenated salicylanilides,benzoic esters, halogenated diphenyl ethers, and mixtures thereof. Aparticularly suitable nonionic antibacterial agent is a diphenyl ethersuch as 2,4,4′-trichloro-2′-hydroxydiphenyl ether (Triclosan) and2,2′-dihydroxy-5,5′-dibromodiphenyl ether. Antifungal agents can be ofthe allylamines type which includes, but is not limited to, amorolfin,butenafine, naftifine and terbinafine.

Antiviral Agents

Antivirals are used for specific viruses and are typically harmless tothe host. Many available drugs available are created to treat infectionsby retroviruses such as HIV. Important antiretroviral drugs include theclass of protease inhibitors. Herpes viruses, best known for causingcold sores and genital herpes, are usually treated with the nucleosideanalogue acyclovir. Viral hepatitis is caused by five unrelatedhepatotropic viruses and is also commonly treated with antiviral drugsdepending on the type of infection. Influenza A and B viruses areimportant targets for the development new influenza treatments toovercome the resistance to existing neuraminidase inhibitors such asoseltamivir.

Antiparasitic Agents

Antiparasitics are a class of medications which are indicated for thetreatment of infection by parasites, such as nematodes, cestodes,trematodes, infectious protozoa, and amoebae.

Antimicrobial Coatings

A surface can be made antimicrobial by providing a coating containing anantibacterial agent that inhibits or diminishes the ability of amicroorganism to grow on the surface of a material. Surfacecontamination has become recognized as a health risk in various settingsincluding clinical, industrial and home. Antimicrobial coatings havebeen commonly used in the healthcare industry for sterilizing medicaldevices to prevent hospital-associated infections. Medical devices,surgical instruments, tubing, suture, tape, bandaging, linens andclothing provide a potential environment for many bacteria, fungi, andviruses to grow when in contact with the human body which allows for thetransmission of infectious disease. Likewise, implantable devices suchas pacemakers and subcutaneous rods provide environments for microbialgrowth, and would pose less risk of infection if treated with anantimicrobial coating. Antimicrobial surfaces can be functionalized in avariety of different processes. A coating may be applied to a surfacethat has a chemical compound which is toxic to a microorganism. Othersurfaces may be functionalized by attaching a polymer or polypeptide toits surface. In other cases, it is advantageous to have an encapsulatedaqueous solution of a peptide-based or peptide analog-basedantimicrobial agent embedded in a surface coating.

Anticoagulants and Antithrombotics

Certain medical procedures expose patients to life-threatening bloodclots. Patients often receive anticoagulant drugs that reduce or preventthe blood from clotting. Anticoagulant therapy is regularly administeredand is a useful aid in the recovery of most patients. However, the useof anticoagulants increases the risk of bleeding, while preserving asufficient supply of blood. Peptide-based and peptide analog-based drugshave exhibited promise in the maintenance of the antithrombosis/bleedingbalance in the patient. This is of value in procedures such aspercutaneous coronary intervention and coronary arterial bypass graft.

Anticoagulants include, but are not limited to, coumadins, heparins andits derivatives, low molecular weight heparins, syntheticpentasaccharide inhibitors such as fondaparinux and Idraparinux, directfactor Xa inhibitors, such as rivaroxaban and apixaban and directthrombin inhibitors including, but not limited to, hirudin, lepirudin,bivalirudin, argatroban and dabigatran.

Anticancer Agents

Therapeutic agents can also be anticancer agents, which include, but arenot limited to, alkylating agents such as mechlorethamine,cyclophosphamide, chlorambucil, and ifosfamide. Anticancer agents can beantimetabolites or plant alkaloids or terpenoids which include, but arenot limited to, vincristine, vinblastine, vinorelbine and vindesine.Anticancer agents can include podophyllotoxin or taxanes. Anticanceragents can be topoisomerase inhibitors including, but not limited to,the camptothecins irinotecan and topotecan, amsacrine, etoposide,etoposide phosphate, and teniposide. Anticancer agents can be cytotoxicantibiotics that include, but are not limited to, actinomycin,anthracyclines, doxorubicin, daunorubicin, valrubicin, idarubicin,epirubicin, bleomycin, plicamycin, and mitomycin. Antiprotozoals caninclude, but are not limited to, eflornithine, furazolidone,melarsoprol, metronidazole, ornidazole, paromomycin sulfate,pentamidine, pyrimethamine and tinidazole.

Anti-Inflammatory Agents

Anti-inflammatory agents include, but are not limited to, flucinoloneand hydrocortisone, ketorolac, flurbiprofen, ibuprofen, naproxen,indomethacin, diclofenac, etodolac, indomethacin, sulindac, tolmetin,ketoprofen, fenoprofen, piroxicam, nabumetone, aspirin, diflunisal,meclofenamate, mefenamic acid, oxyphenbutazone and phenylbutazone.

Antiplaque Agents

Antiplaque (e.g., plaque disrupting) agents can be a form of therapeuticagent. Suitable antiplaque agents include, without limitation,glucoamylase and glucose oxidase.

Desensitizing Agents

Desensitizing, or tooth sensitivity-protecting agents, can betherapeutic agents. Desensitizing agents include, without limitation,potassium salts such as potassium citrate, potassium tartrate, potassiumchloride, potassium sulfate and potassium nitrate or sodium nitrate.

Systemic Analgesics

Systemic analgesics include, but are not limited to, aspirin, codeine,acetaminophen, sodium salicylate and triethanolamine salicylate.

Drug Delivery and Indications

Subcutaneous Drug Delivery

Biodegradable polymeric systems represent a promising method fordelivering many therapeutic agents, including, for example, peptides andpeptide analogs. Some polymers undergo a sol-gel transition onceadministered. A gel can form in situ in response to one or a combinationof stimuli including UV irradiation, pH change, temperature change, andsolvent exchange. Some polymeric systems have several advantages overconventional methods, including ease of manufacturing, ease ofadministration, and biodegradability. There have been challenges,however, to control the release profiles of the incorporated therapeuticagents. It would be advantageous to be able to store a peptide-based orpeptide analog-based drug in a microencapsulated buffer solution,wherein the semipermeable microcapsule could control the rate of releaseof the therapeutic agent. The microcapsule can be made to biodegradealong with the polymer system in which it is embedded.

Controlled Release

The microcapsules and products of the invention can be designed to havedifferent time release profiles, in such a way as to permit controlledtime release, also known as sustained release or long-term release, ofthe therapeutic agents or constituents to effectuate various desiredresults. Thus, various additives or components of the invention can bereleased at varying periods of time from each microcapsule. A pluralityof microcapsules that contain different concentrations can achieve, ineffect, a targeted therapeutic agent release profile.

Furthermore, the ratios of release times in different types ofmicrocapsules can be incorporated into the design of a single product tooptimize the permeability and concentration of the specific biologicallyactive additives or constituents of each type of microcapsule in theproduct relative to one another and relative to the environment wherethey are required. Such design enables a product that can deliver anumber of buffered therapeutic agents over a controlled time to aparticular targeted site without the need for numerous administrationsof a product to a patient, thereby minimizing the treatment regimen ofthe patient.

For example, a dental product can provide an antibacterial treatment toa targeted tooth surface in the oral cavity. For example, fillings,sealants and cements can be designed to contain the controlled releasemicrocapsules of the invention and to release buffered antimicrobialagents over time to the area of contact with the tooth or toothmaterial. Sustained release dosage forms of dental products avoid thenecessity of frequently administering an active while at the same timeachieving a desired level of anticaries activity in the oral cavity.

One method for controlling release is selecting a shell polymer havingdesired permeability. Another method to control permeability and releaseprofile is to control the thickness of the shell layer during synthesisof the microcapsules as described herein.

The concentration of agents within the microcapsule can also be variedto effect permeability. In a preferred embodiment, buffered therapeuticagents and combinations thereof are incorporated in the microcapsules.Each microcapsule is synthesized with a buffered aqueous solution of atherapeutic agent with a specific targeted range of concentrations.

For example, tablets comprising the microcapsules of the invention canbe produced, wherein the therapeutic agent is not all immediatelyabsorbed, but instead is released gradually and continuously over aperiod of time from administration. For prolonged shelf life andsubstantive effects, the therapeutic agent is preferentially stored in abuffered solution.

In the oral cavity, an instant release or “burst” release of themicrocapsules can result upon mechanical agitation of the teeth, such asby use of toothbrushes or dental floss in the oral cavity; by regularchewing, grinding, gritting, clenching or clamping of the teeth or gums;by tongue motion or pressure of the tongue; or by swishing or garglingof liquids by the jaw muscles and motion of other muscles within theinternal orifices of the mouth. Both semipermeable microcapsules and theimpermeable microcapsules of the invention, as described above, can beincorporated into products designed for burst release effect.

Pharmaceutical Compositions

The compositions of this invention can be in the form of pharmaceuticalcomposition comprising the therapeutic agent-containing microcapsulesand a pharmaceutically acceptable carrier. Pharmaceutically acceptablecarriers are well known to those skilled in the art and include, but arenot limited to, 0.01-0.1 M and preferably 0.05 M phosphate buffer or0.8% saline. Additionally, such pharmaceutically acceptable carriers canbe aqueous or nonaqueous solutions, suspensions, and emulsions. Examplesof nonaqueous solvents are propylene glycol, polyethylene glycol,vegetable oils such as olive oil, and injectable organic esters such asethyl oleate. Aqueous carriers include water, alcoholic/aqueoussolutions, emulsions and suspensions, including saline and bufferedmedia. Parenteral vehicles include sodium chloride solution, Ringer'sdextrose, dextrose and sodium chloride, lactated Ringer's and fixedoils. Intravenous vehicles include fluid and nutrient replenishers,electrolyte replenishers such as Ringer's dextrose, those based onRinger's dextrose, and the like. Preservatives and other additives mayalso be present, such as, for example, antimicrobials, antioxidants,chelating agents, inert gases, and the like.

Bone Restorative Materials

The compositions of the invention are useful in a variety of bonerestorative or regeneration products. There is a need for new materialsthat can stimulate the body's own regenerative mechanisms and healtissues. Porous templates that act as scaffolds are thought to berequired for three-dimensional bone tissue growth. Bone growth factorshave high potential to stimulate bone-forming cells to produce new bone,they are degradable in the body and they bond to bone. Bone restorativeor regeneration products may include bone growth factors that could bestored longer or have more substantive effects if stored in a bufferedenvironment within the microcapsule.

Specific dental products comprising the microcapsules of the inventioninclude dental gels, pastes, rinses, dentifrices, whitening products,breath fresheners, artificial saliva systems, varnishes, desensitizersand other dental products well known in the dental art. Dentalrestorative materials include, but are not limited to, composite andother solid phase filling materials, adhesives and cements, temporaryrestorative materials, coatings on implants for the induction of bonegrowth. Various embodiments of the invention include over-the-counterapplications such as toothpastes, bleaching agents, varnishes, sealants,sealers, resin restorative materials, glass ionomers (including resinmodified glass ionomers), bioactive glass, compomer restorativematerials, giomer restorative materials, oral rinses, any topicalpreventive or remineralizing agents (liquids, gel mousses, pastes), anyrinse including antimicrobial agents, professionally applied andover-the-counter “paint-on” liquids gels, varnishes, sealers, indirectlaboratory materials including laboratory resins, denture teeth, denturebase materials, dental cements, root canal fillers and sealers,materials used for bone grafting, bone cements, dental implant tissuegrowth materials, endodontic root filling materials (i.e. apicoectomymaterials sometimes called retro-fill materials), pulp cappingmaterials, temporary restorative filling materials, prophy paste,periodontal scaling gels, air abrasion powders for prophylaxis,orthodontic cements, oral surgery extraction socket dressing, cadaverbone, and other bone substitutes.

Embodiments of dental products also include products that can dissolvein the oral cavity upon contact with saliva as a result of enzymaticactivity in the oral cavity, such as dissolvable whitening strips. Otherproducts incorporating the microcapsules that have a targeted effect onteeth include chewing gums, candies, lozenges, capsules, tablets andvarious food items.

In an embodiment where the microencapsulated dental compositions of thespecific buffered solutions of antimicrobial agents of the inventionallow incorporation of antimicrobial agents into a matrix ofpolymerizable composites and other solid filling dental restorativematerials such as glass ionomer cements, the incorporation of themicroencapsulated antimicrobial agents provides a source of bioactivefilling materials and adhesives. The use of semipermeable microcapsulesallows the material to release these antimicrobial agents at theinterface between the tooth structure and the restorative fillingmaterial or adhesive. This interface is particularly vulnerable tobacterial ingress, attack and subsequent secondary caries development.The presence of fluid within this interface can signal possiblemicroleakage at the restorative interface but also allow activation ofthe material to release the antimicrobial agents. Embodiments of themicrocapsules of the invention can be designed to release theantimicrobial agents when under mechanical stress at the opening of aspace at the tooth/filling interface.

Dental composition embodiments of the invention also comprise a solidphase, such as composites, which offer multiple advantages. Presently,it is unknown to add antimicrobial agents into a resin composite andprovide activity of the antimicrobial agents because the antimicrobialagents are likely incorporated or entombed within the resin or plasticinsoluble matrix.

Delivery and Indications for Subcutaneous and Topical Therapeutic Agents

Related Therapeutic Products for Topical Use on Hair, Nails, Skin andOther

Epithelial Tissue

There are numerous therapeutic agents that can be delivered to theepithelial tissues of a subject via products such as moisturizers,creams, lotions, foams and gels. Such agents include the followingnonlimiting examples: antibacterials such as Bactroban or Cleocin;Anthralin (Drithocreme, Micanol and others) for psoriasis; antifungalagents such as Lamisil, Lotrimin and Nizoral for skin conditions such asringworm and athlete's foot; benzoyl peroxide creams for treating acne;coal tar for treating conditions such as seborrheic dermatitis (usuallyvia shampoos) or psoriasis; corticosteroids for treating skin conditionsincluding eczema via foams, lotions, ointments and creams; retinoids(such as Retin-A and Tazorac) are gels or creams for treating acne;salicylic acid in lotions, gels, soaps, shampoos, and patches, is usedto treat acne and warts; antiviral agents such as Valtrex, acyclovir,and Famvir are useful for treating herpes; corticosteroids such asprednisone, and immunosuppressants, such as azathioprine andmethotrexate, are useful in treating inflammatory diseases such aseczema and psoriasis; and biologics such as Enbrel, Humira, Remicade,Stelara and Amevive are useful for treating psoriasis as well.

Shampoo

A shampoo is made by combining a surfactant (typically sodium laurylsulfate and/or sodium laureth sulfate) with a cosurfactant (usuallycocamidopropyl betaine) in water to form a thick, viscous liquid. Othershampoo components include salt (e.g., sodium chloride), preservativesand fragrances. Further components are usually added to achieve thefollowing properties: pleasing foam; easy rinsing; minimal skin and eyeirritation; thick and creamy feel; good fragrance; low toxicity;biodegradability; and proper pH.

The following are common shampoo ingredients: glycol distearate;silicone; ammonium chloride; ammonium lauryl sulfate; glycol; sodiumlauroamphoacetate (cleanser and counterirritant); polysorbate 20(abbreviated as PEG(20), penetrant); polysorbate 80 (abbreviated asPEG(80), emulsifier); PEG-150 distearate (thickener); citric acid(antioxidant, pH adjuster and preservative); quaternium-15(preservative); polyquaternium-10 (conditioning agent); Di-PPG-2myreth-10 adipate (emollient); and methylisothiazolinone (MIT,preservative).

Hair Conditioner

Hair conditioners contain many types of ingredients. The following areexamples: moisturizers (e.g., humectants); reconstructors (e.g.,hydrolyzed protein); acidifiers; detanglers (e.g., polymers); thermalprotectors (e.g., heat-absorbing polymers); glossers (e.g., siliconessuch as dimethicone or cyclomethicone); oils (i.e., essential fattyacids); surfactants (cationic); lubricants (e.g., fatty alcohols,panthenol, and dimethicone); sequestrants, for better function in hardwater; antistatic agents; and preservatives. Conditioners include, forexample, the pack conditioners, leave-in conditioners, ordinaryconditioners, and hold conditioners.

Hair Gel

Hair gels, which contain cationic polymers, are hairstyling productsthat are used to stiffen hair into a particular hairstyle.

Moisturizers

Moisturizers increase the skin's hydration (water content) by reducingevaporation. Naturally occurring skin lipids and sterols as well asartificial or natural oils, humectants, emollients, and lubricants,etc., may be part of the composition of commercial skin moisturizers.

Moisturizers for preserving normal skin contain, e.g., lightweight oils,such as cetyl alcohol, or silicone-derived ingredients, such ascyclomethicone. Moisturizers for treating dry skin contain ingredientssuch as antioxidants, grape seed oil or dimethicone, and petrolatum (forvery dry skin). Moisturizers for treating the effects of aging contain,e.g., petrolatum, antioxidants and alpha hydroxy acids.

Nail Polish

Nail polishes typically contain phthalates (e.g., dibutylphthalate(DBP), dimethylphthalate (DMP), diethylphthalate (DEP)), and toluene.

Additives

As used herein, the terms “additive” and “therapeutic agent” are notmutually exclusive. Although many additives have no known therapeuticrole, some additives do have known therapeutic benefits.

In some embodiments, where oral application is desired, a sweetener isemployed in products that incorporate the microcapsule compositions ofthe present invention. Sweeteners among those useful herein include,without limitation, orally acceptable natural or artificial, nutritiveor nonnutritive sweeteners. Such sweeteners include, without limitation,dextrose, polydextrose, sucrose, maltose, dextrin, dried invert sugar,mannose, xylose, ribose, fructose, levulose, galactose, corn syrup(including high fructose corn syrup and corn syrup solids), partiallyhydrolyzed starch, hydrogenated starch hydrolysate, sorbitol, mannitol,xylitol, maltitol, isomalt, aspartame, neotame, saccharin and saltsthereof, sucralose, dipeptidebased intense sweeteners, cyclamates,dihydrochalcones, and mixtures thereof. One or more sweeteners can bepresent in a total amount depending strongly on the particularsweetener(s) selected,

Products incorporating the compositions of the present invention, whereoral application is desired, optionally comprise a flavoring agent.Flavoring agents among those useful herein include any material ormixture of materials operable to enhance the taste of the composition.Any orally acceptable natural or synthetic flavorant can be used, suchas flavoring oils, flavoring aldehydes, esters, alcohols, similarmaterials, and combinations thereof. Flavoring agents include vanillin,sage, marjoram, parsley oil, spearmint oil, cinnamon oil, oil ofwintergreen (methyl salicylate), peppermint oil, clove oil, bay oil,anise oil, eucalyptus oil, citrus oils, fruit oils and essencesincluding those derived from lemon, orange, lime, grapefruit, apricot,banana, grape, apple, strawberry, cherry, pineapple, etc., bean- andnut-derived flavors such as coffee, cocoa, cola, peanut, almond, etc.,adsorbed and encapsulated flavorants, and mixtures thereof. Alsoencompassed within flavoring agents are ingredients that providefragrance and/or other sensory effect in the mouth, including cooling orwarming effects. Such ingredients include, for example, menthol, menthylacetate, menthyl lactate, camphor, eucalyptus oil, eucalyptol, anethole,eugenol, cassia, oxanone, a-irisone, propenyl guaiethol, thymol,linalool, benzaldehyde, cinnamaldehyde, N-ethyl-p-menthan-3-carboxamine,N,2,3trimethyl-2-isopropylbutanamide, 3-1-menthoxypropane-1,2-diol,cinnamaldehyde glycerol acetal (CGA), methone glycerol acetal (MGA), andmixtures thereof.

In some embodiments of the invention, the therapeutic agent is a“systemic active” which is used to treat or prevent a disorder which, inwhole or in part, is not a disorder of the oral cavity. In variousembodiments, the active is an “oral care active” used to treat orprevent a disorder or provide a cosmetic benefit within the oral cavity(e.g., to the teeth, gingiva or other hard or soft tissue of the oralcavity). Oral care actives among those useful in the dental compositionsherein include anticaries agents, tartar control agents, periodontalactives, abrasives, breath freshening agents, malodour control agents,tooth desensitizers, salivary stimulants, and combinations thereof. Itis understood that while general attributes of each of the abovecategories of therapeutic agent may differ, there may some commonattributes and any given material may serve multiple purposes within twoor more of such categories of agents. These therapeutic agents maypreferably be stored in a buffered solution to improve shelf life andprovide more substantive effects.

Products comprising the compositions of the present invention optionallycomprise an antioxidant. Any topically acceptable antioxidant can beused, including, but not limited to: butylated hydroxyanisole (BHA),butylated hydroxytoluene (BHT), vitamin A, carotenoids, vitamin E,flavonoids, polyphenols, ascorbic acid, herbal antioxidants,chlorophyll, melatonin, and mixtures thereof. Products comprising anantioxidant may preferably store the composition containing theantioxidant in a buffered solution to improve shelf life and providemore substantive effects.

Products containing compositions of the present invention optionallycomprise an orally acceptable zinc ion source useful, for example, as anantimicrobial, anticalculus or breath-freshening agent. One or more suchsources can be present. Suitable zinc ion sources include, withoutlimitation, zinc acetate, zinc citrate, zinc gluconate, zinc glycinate,zinc oxide, zinc sulfate, sodium zinc citrate and the like. The productsof the invention can optionally comprise a suitable pH-adjusting agent,including, but not limited to, sodium hydroxide, potassium hydroxide,and ammonium, for controlling the stability and shelf life of a dentalproduct.

Products such as food materials incorporating the compositions of thepresent invention optionally comprise a nutrient. Suitable nutrientsinclude vitamins, minerals, amino acids, and mixtures thereof. Vitaminsinclude Vitamins C and D, thiamine, riboflavin, calcium pantothenate,niacin, folic acid, nicotinamide, pyridoxine, cyanocobalamin,para-aminobenzoic acid, bioflavonoids, and mixtures thereof. Nutritionalsupplements include amino acids (such as L-tryptophane, L-lysine,methionine, threonine, levocarnitine and L-carnitine), lipotropics (suchas choline, inositol, betaine, and linoleic acid), fish oil (includingcomponents thereof such as omega-3 (N-3) polyunsaturated fatty acids,eicosapentaenoic acid and docosahexaenoic acid), coenzyme Q10, andmixtures thereof. Products comprising a nutrient or nutritionalsupplement may preferably store the composition containing the nutrientor nutritional supplement in a buffered solution to improve shelf lifeand provide more substantive effects.

In another embodiment of the invention, the products comprising themicrocapsules may further include an antibacterial agent for releaseonto the bone tissue or dental surface. A wide variety of antimicrobialactive compounds may be employed. These actives may generally beclassified as halogenated hydrocarbons, quaternary ammonium salts andsulfur compounds. Halogenated hydrocarbons include halogenatedderivatives of salicylanilides, carbanilides, bisphenols, diphenylethers, anilides of thiophene carboxylic acids and chlorhexidines.Quaternary ammonium compounds include alkyl ammonium, pyridinum, andisoquinolinium salts. Sulfur active compounds include thiuram sulfidesand dithiocarbamates. Products comprising an antibacterial agent maypreferably store the composition containing the antibacterial agent in abuffered solution to improve shelf life and provide more substantiveeffects.

In various embodiments of the dental products of the present invention,the dental product comprises an adhesive or adhesion-enhancing agentwhich serves multiple functions, including enhancing adherence of acomposition to the surface of the tooth to be remineralized or whitened.The adhesives are optimized for adhering to the teeth, resistingadherence to non-tooth oral surfaces such as the lips, gingival or othermucosal surfaces, and remaining attached to the teeth for an extendedtime. Optimization of such aspects may be achieved from varying thephysical and chemical properties of a single adhesive or combiningdifferent adhesives. In certain embodiments of the present invention,the adhesive polymers in the product are those in which a dentalparticulate can be dispersed and are well known in the art.

The compositions of this invention can also be incorporated intocandies, lozenges, chewing gums, tablets, capsules or other products.Incorporation of the microcapsules into the product can be achieved by,for example, stirring into a warm gum base or coating the outer surfaceof a gum base, illustrative of which may be jelutong, rubber latex,vinylite resins, and similar compounds desirably with conventionalplasticizers or softeners, sugar, glucose, sorbitol or other sweeteners.It is also contemplated herein that the microcapsule compositions of thepresent invention can be incorporated into a variety of food items.

Each of the instant compositions, products and methods is envisioned forhuman use. It is also understood that although this specification refersspecifically to applications in humans, the invention is also useful forveterinary purposes. Thus, in all aspects, the methods and compositionsof the invention are useful for domestic animals such as cattle, sheep,horses and poultry; and for companion animals such as cats and dogs; aswell as for other animals.

It will be appreciated by persons skilled in the art that the presentinvention is not limited to the specific embodiments that have beenparticularly shown and described hereinabove. Rather the scope of thepresent invention includes combinations of the features described aswell as modifications and variations thereof which would occur to aperson of skill in the art upon reading the foregoing description andwhich are not in the prior art.

EXAMPLES

The following examples set forth the compositions and the synthesismethods of the invention. These experiments demonstrate the feasibilityof using the surfactant-free interfacial polymerization of a reverseemulsion to successfully encapsulate buffered solutions of therapeuticagents to create effective therapeutic compositions.

Example 1

A microcapsule composition of the invention was prepared containing abuffered solution of a peptide-based anitimicrobial agent. Thecomposition was prepared by performing an interfacial polymerization ina stable inverse emulsion of phosphate buffer saline solution of apeptide-based antimicrobial agent in a methyl benzoate continuous phase.Six grams of polyglyceryl-3-polyricinoleate (P3P) was used as theemulsifying agent. The emulsifying agent and 4 grams of a polyurethanepolymer were mixed together. The aqueous buffer solution of thepeptide-based antimicrobial agent (100 mL of 0.1 M) was added to 210 mLof continuous methyl benzoate oil phase under mixing. 0.2 g of ethyleneglycol was subsequently added to the inverse emulsion to complete theinterfacial polymerization of the polyurethane polymer at the interfaceof the dispersed buffered aqueous solution of peptide-basedantimicrobial agent droplet. The average size of the microcapsule wascontrolled by the rate of mixing. Using the method of the invention, thefollowing buffered aqueous solutions of an antimicrobial agent werethereby prepared.

Example 2

A microcapsule composition of the invention was prepared containing abuffered solution of a peptide-based anitimicrobial agent. Thecomposition was prepared by performing an interfacial polymerization ina stable inverse emulsion of phosphate buffer saline solution in amethyl benzoate continuous phase. Six grams ofpolyglyceryl-3-polyricinoleate (P3P) was used as the emulsifying agent.The emulsifying agent and 4 grams of a polyurethane polymer were mixedtogether and added to 210 mL of continuous methyl benzoate oil phaseunder mixing. 0.2 g of ethylene glycol was subsequently added to theinverse emulsion to complete the interfacial polymerization of thepolyurethane polymer at the interface of the dispersed buffered aqueoussolution of a peptide-based antimicrobial agent. The microcapsulescontaining buffer solution were stirred in a solution of a bufferedpeptide-based antimicrobial agent (100 mL of 0.1 M) while being mixedand heated to 37° C. to effectively load the microcapsule with theantimicrobial agent. Using the method of the invention, the followingbuffered aqueous solutions of an antimicrobial agent were therebyprepared.

Example 3

A composition of the invention for cavity varnish with antimicrobialcapabilities was prepared as follows. A standard cavity varnishcontaining rosin, ethanol and thymol (97 wt %) were combined with 3 wt %of a microcapsule containing a 0.1 M buffered aqueous solution of anantimicrobial agent.

Example 4

A composition for toothpaste with antimicrobial capabilities wasprepared comprising a colloidal binding agent, humectants,preservatives, flavoring agents, abrasives, and detergents. 2 wt % of amicrocapsule containing a buffered aqueous solution of a peptide-basedantimicrobial agent was incorporated. The antimicrobial agents will bereleased from the encapsulated buffered solution to improvemineralization of the teeth.

Example 5

A composition for a dental resin composite with remineralization andtherapeutic capabilities was prepared as follows. A resin mixture (16 wt% total) was first made by combining urethane dimethacrylate resin withtriethyleneglycoldimethacrylate (TEGDMA) resin in a 4:1 ratio. Aphotosensitizer (camphoroquinone) was added at 0.7 wt % of the totalcomposition. An accelerator (ethyl-4-dimethylaminobenzoate) was added at3 wt % of the total composition. An inhibitor (4-methoxyphenol) wasadded at 0.05 wt % of the total composition. The resin, photosensitizer,accelerator and inhibitor were combined in a flask and mixed at 50° C.Upon homogenization, the above resin blend was mixed with the followingfillers (84 wt % total): silanated strontium glass 71 wt %, fumed silica10 wt %, microcapsules containing a buffered solution of peptide-basedantimicrobial agents.

What is claimed is:
 1. A topical microcapsule composition comprising anantibacterial agent comprising: a plurality of microcapsules and acarrier, each of said microcapsules comprising at least one polymersubstantially disposed as a semipermeable shell around a bufferedsolution, said microcapsule composition comprising at least a firstsubset of microcapsules, wherein the first subset of microcapsulescomprises within each of said microcapsules the antibacterial agent fortopical use on epithelial tissue; wherein the antibacterial agentpermeates the shell.
 2. The microcapsule composition of claim 1 whereinthe buffered solution is an aqueous solution.
 3. The microcapsulecomposition of claim 1 wherein the antibacterial agent is a naturalantimicrobial agent selected from the group consisting of: beta lactamantibiotics, penicillins cephalosporins, protein synthesis inhibitors,aminoglycosides, macrolides, ketolides, tetracyclines, chloramphenicol,polypeptides, penicillin G, procaine penicillin, benzathine penicillin,penicillin V, cefacetrile, cefadroxil, cephalexin, cefaloglycin,cefalonium, cefaloridine, cefalotin, cefapirin, cefatrizine, cefazaflur,cefazedone, cefazolin, cefradine, cefroxadine, ceftezole, cefaclor,cefonicid, cefprozil, cefuroxime, cefuzonam, cefmetazole, cefotetan,cefoxitin, amikacin, arbekacin, gentamicin, kanamycin, neomycin,netilmicin, paromomycin, rhodostreptomycin, streptomycin, tobramycin,apramycin, azithromycin, clarithromycin, dirithromycin, erythromycin,roxithromycin, telithromycin, telithromycin, cethromycin, solithromycin,spiramycin, ansamycin, oleandomycin, carbomycin, tylosin, tetracycline,chlortetracycline, oxytetracycline, demeclocycline doxycycline,lymecycline, meclocycline, methacycline, minocycline, rolitetracycline,actinomycin, bacitracin, colistin, polymyxin B, sulphonamides,cotrimoxazole, quinolones, sulfamethoxazole, sulfisomidine,sulfacetamide, sulfadoxine, dichlorphenamide, dorzolamide,acetazolamide, bumetanide, chlorthalidone, clopamide, furosemide,hydrochlorothiazide, indapamide, mefruside, metolazone, xipamide,acetazolamide, ethoxzolamide, sultiame, zonisamide, celecoxib,darunavir, probenecid, sulfasalazine, and sumatriptan.
 4. A method offorming a topical microcapsule composition comprising a plurality ofmicrocapsules: said plurality of microcapsules being formed by combininga polymer and at least one buffered aqueous solution comprising atherapeutic agent, by contacting: (a) said buffered solution comprisingthe therapeutic agent, (b) an oil phase, (c) said polymer, and (d) anemulsifying agent, wherein the polymer substantially forms asemipermeable shell around the buffered aqueous solution; and addingsaid plurality of microcapsules to a carrier.
 5. The method of claim 4wherein said plurality of microcapsules comprises a first and a secondplurality of microcapsules, wherein a first plurality of microcapsulesis formed using a first therapeutic agent, and a second plurality ofmicrocapsules is formed using a second therapeutic agent.
 6. The methodof claim 5 wherein the first and second pluralities of microcapsuleshave a different release profile.
 7. The method of claim 4 furthercomprising a diol, an isocyanate, or both, wherein said diol,isocyanate, or both increases the molecular weight of the semipermeableshell.
 8. The method of claim 4 wherein the oil phase is methylbenzoate.
 9. The method of claim 4 wherein the emulsifying agent is apolyglyceryl-3-polyricinoleate.
 10. The method of claim 4 wherein saidtherapeutic agent is selected from the group consisting of:antibacterial agents, antifungal agents, benzoyl peroxide, coal tar,corticosteroids, retinoids, salicylic acid, antiviral agents,immunosuppressants, and biologic agents for treating psoriasis.
 11. Themethod of claim 4 wherein the carrier is in the form of a moisturizer,cream, lotion, foam, gel, shampoo, hair conditioner, hair gel, or a nailpolish.
 12. The method of claim 4 wherein the therapeutic agent is abiologically active additive or a peptide-based or peptide analog-basedantimicrobial agent.
 13. The method of claim 4 further comprising anatural oil, a synthetic oil, flavoring oils, flavoring aldehydes,esters, alcohols, vanillin, sage, marjoram, parsley oil, spearmint oil,cinnamon oil, oil of wintergreen, peppermint oil, clove oil, bay oil,anise oil, eucalyptus oil, citrus oil, lemon oil, orange oil, lime oil,grapefruit oil, apricot oil, banana oil, grape oil, apple oil,strawberry oil, cherry oil, pineapple oil, bean-derived oil, nut-derivedoil, cocoa oil, cola oil, peanut oil, almond oil, and combinationsthereof.
 14. The method of claim 4 wherein the therapeutic agent isselected from the group consisting of: menthol, menthyl acetate, menthyllactate, camphor, eucalyptus oil, eucalyptol, anethole, eugenol, cassia,oxanone, a-irisone, propenyl guaiethol, thymol, linalool, benzaldehyde,cinnamaldehyde, N-ethyl-p-menthan-3-carboxamine,N,2,3trimethyl-2-isopropylbutanamide, 3-1-menthoxypropane-1,2-diol,cinnamaldehyde glycerol acetal (CGA), methone glycerol acetal (MGA), andcombinations thereof.
 15. The method of claim 4 wherein the therapeuticagent is selected from the group consisting of: butylated hydroxyanisole(BHA), butylated hydroxytoluene (BHT), vitamin A, carotenoids, vitaminE, flavonoids, polyphenols, ascorbic acid, herbal antioxidants,chlorophyll, melatonin, and combinations thereof.
 16. The method ofclaim 4 wherein the therapeutic agent is selected from the groupconsisting of: salicylanilides, carbanilides, bisphenols, diphenylethers, anilides of thiophene carboxylic acids, chlorhexidines, alkylammonium, pyridinium, isoquinolinium salts, thiuram sulfides,dithiocarbamates, and combinations thereof.
 17. The method of claim 4wherein the therapeutic agent is selected from the group consisting of:a natural antimicrobial agent selected from the group consisting of:beta lactam antibiotics, penicillins, cephalosporins, protein synthesisinhibitors, aminoglycosides, macrolides, ketolides, tetracyclines,chloramphenicol, polypeptides, penicillin G, procaine penicillin,benzathine penicillin, penicillin V, cefacetrile, cefadroxil,cephalexin, cefaloglycin, cefalonium, cefaloridine, cefalotin,cefapirin, cefatrizine, cefazaflur, cefazedone, cefazolin, cefradine,cefroxadine, ceftezole, cefaclor, cefonicid, cefprozil, cefuroxime,cefuzonam, cefmetazole, cefotetan, cefoxitin, amikacin, arbekacin,gentamicin, kanamycin, neomycin, netilmicin, paromomycin,rhodostreptomycin, streptomycin, tobramycin, apramycin, azithromycin,clarithromycin, dirithromycin, erythromycin, roxithromycin,telithromycin, telithromycin, cethromycin, solithromycin, spiramycin,ansamycin, oleandomycin, carbomycin, tylosin, tetracycline,chlortetracycline, oxytetracycline, demeclocycline, doxycycline,lymecycline, meclocycline, methacycline, minocycline, rolitetracycline,actinomycin, bacitracin, colistin, polymyxin B, sulphonamides,cotrimoxazole, quinolones, antivirals, antifungals, anticancer drugs,antimalarials, antituberculosis drugs, antileprotics, antiprotozoals,sulfamethoxazole, sulfisomidine, sulfacetamide, sulfadoxine,dichlorphenamide, dorzolamide, acetazolamide, bumetanide,chlorthalidone, clopamide, furosemide, hydrochlorothiazide, indapamide,mefruside, metolazone, xipamide, acetazolamide, ethoxzolamide, sultiame,zonisamide, celecoxib, darunavir, probenecid, sulfasalazine, andsumatriptan.